Summary: Since both humoral and cellular and immune responses may be important against AIDS, two HIV-1 multigenic plasmid DNAs were constructed containing the gag and env genes from pNL4-3 DNA under the regulation of the human CMV and murine endogenous AKV promoters. Development of humoral and T cell proliferative immune responses were analyzed by direct DNA injection of rhesus and pig-tailed macaques. Similar levels of T-cell proliferative responses were generated with both DNAs in rhesus macaques whereas the humoral responses in rhesus and pig-tailed macaques correlated directly with the promoter strength of the vaccine DNA. Long-term, boostable antibodies to Gag and Env were generated using less amount and fewer injections of CMV-DNA than with AKV-DNA. Tetanus toxoid injection was used to demonstrate ability of the animals to respond similarly to a known antigen. To evaluate protection against HIV-1, DNA vaccinated pig-tailed macaques were boosted with a single injection of oligomeric gp160 (to enhance the env response). Evaluation of the Env antibody responses at 4 weeks post-boost indicated increased antibody titers to gp120 and gp41 in the HIV-1 DNA vaccinated animal as compared with the control animals. Pig-tailed macaques were challenged with HIV-1/LAI. The efficacy of the vaccine DNAs was analyzed by evaluating plasma viral load and detection of sequences in the PBMCs. The results indicated vaccine efficacy correlated with promoter strength.